去泛素化酶Otulin基因心内膜敲除小鼠模型的构建及表型初步分析

Phenotypes of a mouse model of specific deubiquitinase Otulin gene knockout in the endocardium

目的构建心内膜条件性敲除去泛素化酶Otulin基因小鼠模型,初步分析该模型小鼠的表型。方法Cre?loxP重组技术构建心内膜条件性Otulin基因敲除小鼠模型;PCR技术明确Otulin基因敲除小鼠的基因型;统计分析敲除小鼠出生情况并观察其生长发育状态:超声监测小鼠心脏功能,全自动血液分析仪测定血液中细胞组分变化,苏木素?伊红(HE)染色评价小鼠各器官组织的病理改变。结果心内膜特异性Otulin基因敲除小鼠模型成功构建;该条件性敲除小鼠可正常交配繁殖,子代基因型比例遵循孟德尔遗传定律;与同窝对照小鼠相比,该条件性敲除小鼠体型较小,体重减轻,脾明显肿大,血液中的炎性细胞数量显著增加,并且心脏组织三尖瓣缺损,心功能衰减。结论成功构建心内膜特异性Otulin基因敲除小鼠模型,该模型小鼠心脏功能受损且有明显的炎症反应。

Objective To construct a mouse model with conditional knockout of deubiquitinase Otulin gene in the endocardium and to analyze the phenotype of the model.Methods A model of mice with endocardium-specific conditional Otulin gene knockout was constructed with the Cre-loxP technique.The genotypes of Otulin gene knockout mice were determined by using polymerase chain reaction(PCR).Data on the reproduction of the knockout mice was statistically analyzed and the growth and development of the offspring were observed.The Cardiac function of mice was monitored by ultrasound and the changes of cellular components in blood were measured by a fully automatic hematology analyzer.Hematoxylin-eosin(HE)staining was used to analyze the pathological changes in all the tissues and organs of the offspring.Results An endocardium-specific Otulin gene knockout mouse model was established.The conditional knockout mice could grow and reproduce normally and the percentage of genotypes of their offspring conformed to Mende's law.Compared with littermate control mice,the conditional knockout mice had smaller body size,lighter weight,significantly enlarged spleens and spiking inflammatory cells in blood.In addition,the tricuspid valve defect and cardiac function were attenuated.Conclusion A mouse model of endocardium-specific Otulin gene knockout has been constructed.These mice show impaired cardiac function and obvious inflammatory response.